HER2 Status in Premalignant, Early, and Advanced Neoplastic Lesions of the Stomach.

OBJECTIVES: HER2 expression in gastric cancer (GC) has received attention as a potential target for therapy with Trastuzumab. We reviewed the current knowledge on HER2 status in premalignant gastric lesions and in early (EGC) and advanced (AGC) GC to discuss the possible pathogenetic and prognostic roles of HER2 overexpression in GC. RESULTS: HER2 overexpression was documented in gastric low-grade (LG) and high-grade intraepithelial neoplasia (HG-IEN), with higher frequency in gastric type dysplasia. HER2 overexpression was significantly associated with disease recurrence and poor prognosis in EGC representing an independent risk factor for lymph node metastases. HER2 overexpression was more frequent in AGC characterized by high grade, advanced stage, and high Ki-67 labeling index. The discordance in HER2 status was evidenced between primitive GC and synchronous or metachronous metastases. CONCLUSIONS: HER2 overexpression in premalignant gastric lesions suggests its potential involvement in the early steps of gastric carcinogenesis. The assessment of HER2 status in EGC may be helpful for the identification of patients who are at low risk for developing nodal metastases. Finally, the possible discordance in HER2 status between primary GC and its synchronous metastases support routine assessment of HER2 both in the primary GC and in its metastatic lesions.

Phenotypical and genotypical expression of Wolfram syndrome in 12 patients from a Sicilian district where this syndrome might not be so infrequent as generally expected.

BACKGROUND: Since the original description, there have been only few epidemiological studies of Wolfram syndrome (WS). AIM: Aims of the present paper are to ascertain WS prevalence and expression in a district of North-eastern Sicily, i.e. a geographic area where consanguineous unions are not very unusual. MATERIALS AND METHODS: Prevalence rates of WS in the Messina district were calculated by taking into consideration both the total population (653,737) and the populations included within the 0-30 year age range (202,681). We estimated the relative prevalence of WS among patients with youth-onset insulin-dependent diabetes mellitus (DM) who are currently aged under 30 years (256). RESULTS: Global WS prevalence in our district is 1:54,478, whereas prevalence among individuals under 30 is 1:16,890 and relative prevalence among patients with juvenile-onset insulin-dependent DM is 1:22.3. When compared with the patients with insulin-dependent DM of Messina district, WS patients did not exhibit significant differences in terms of biochemical features at DM onset, whereas age at DM diagnosis was significantly earlier in WS group. CONCLUSIONS: (a) WS prevalence is not so infrequent as generally expected; (b) in our series, DM presented before 10 years in 11/12 patients and ten cases have already developed all the four peculiar manifestations of WS by 26 years; (c) 9/12 patients exhibited a homozygous frameshift/truncation mutation (Y454_L459del_fsX454), which is the one most frequently found also in patients from other Italian regions; (d) age at DM diagnosis was significantly earlier in WS group than in the patients with insulin-dependent DM of Messina district.

Neutrophil-rich gastric carcinomas: light and electron microscopic study of 9 cases with particular reference to neutrophil apoptosis.

The authors report 9 cases of gastric carcinomas characterized by a prominent neutrophilic infiltration of the stroma. These tumors (8 of intestinal type, 1 of diffuse type) showed a pushing growth pattern. Metastatic involvement of regional lymph nodes was seen in 5 cases. The metastatic foci were associated with heavy neutrophilia as well. There was no histologic evidence of Helicobacter pylori infection, whereas various degrees of multifocal intestinal metaplasia were present in the background mucosa. Based on histologic and histochemical results, there were no apparent causes due to other infectious agents responsible for the neutrophil-rich gastric carcinomas. Some of intraepithelial and stromal neutrophils exhibited apoptotic changes, such as chromatin condensation and cell shrinkage, and were TUNEL-positive. Electron microscopy disclosed apoptotic neutrophils in cytoplasmic vacuoles of tumor cells, a finding suggestive of neutrophil-tumor cell phagocytosis (cannibalism). Different stages of neutrophil apoptosis were also shown by electron microscopy and the ultrastructural findings were compared to those described in experimental models, both in vivo and in vitro.

Foveolar cells phagocytose apoptotic neutrophils in chronic active Helicobacter pylori gastritis.

The recognition and removal of apoptotic inflammatory cells by tissue macrophages and non-professional phagocytes, in a process called efferocytosis, is required for resolution of inflammation and is actively anti-inflammatory. We have previously demonstrated phagocytosis of apoptotic neutrophils by tumor cells in human gastric carcinoma, but to date, there have been no studies investigating this process in chronic active Helicobacter pylori gastritis. Biopsy specimens from 28 subjects with or without H. pylori infection and active inflammation were examined and graded according to the updated Sydney system. Light microscopy, electron microscopy, and Terminal Deoxynucleotidyltransferase-Mediated UTP End Labeling staining were used to identify apoptosis. H. pylori infection was detected by histology and by molecular assay in 16 out of 28 cases. DNA from paraffin-embedded gastric biopsies was amplified using primers specific for cagA, for the cag "empty site" as well as for the s and m alleles of vacA. The more virulent cagA-positive strains were found in five out of nine patients with chronic active gastritis. The vacA s1/m1 and s2/m1 genotypes were more common in nine patients with chronic active gastritis, while the vacA s2/m2 genotype was more frequent in seven patients with chronic inactive gastritis. Apoptotic neutrophils were also detected within the cytoplasmic vacuoles of the foveolar cells of nine cases with chronic active gastritis. Transmission electron micrographs revealed further apoptotic neutrophils within spacious phagosomes of foveolar cells in a similar manner to those described in late-phase efferocytosis both in vivo and in vitro. These new observations expand the morphological spectrum of gastritis in patients infected with more virulent H. pylori strains, compatible with an anti-inflammatory role for the gastric epithelial cells in their removal of apoptotic neutrophils during active chronic gastritis.

Serum IL-23 in asthmatic children.

Asthma is characterized by airway inflammation that is controlled by a complex cytokine network. The Th1/Th2 imbalance has been well documented in the pathogenesis of allergic asthma. Recently, Th17 cells and regulatory T (Treg) cells have been found to participate in the pathogenesis of allergic asthma. This study aimed at verifying whether anti-inflammatory treatment could change serum IL-4, IL-10 and IL-23 in asthmatic children. Globally, 78 children (40 males and 38 females, median age 9.3 +- 3.7 years), with asthma and monosensitized to house dust mites, were evaluated. Lung function (such as FEV1) and serum IL-4, IL-10 and IL-23 levels were measured at baseline (T0), after 4 weeks (T1) and after 12 weeks (T2) of inhaled corticosteroid (ICS) treatment. The control group consisted of 40 healthy children (22 males and 18 females) age matched. At baseline, IL-4 and IL-23 levels were higher in severe asthmatics than in control group (p less than 0.001), while serum IL-10 levels were significantly lower in group of asthmatic children as compared to healthy control group (p less than 0.001). At T2, IL-4 and IL-23 significantly diminished (p less than 0.001), while IL-10 significantly increased. There was significant relationship between FEV1 and IL-4, IL-10 and IL-23 at T0 (r=-0.784; r=-0.735 and r=-0.787, respectively). Moreover, there were correlations between FEV1 and IL-4, IL-10 and IL-23 in patients at T1 (r=-0.563; r=-0.539 and r=-0.583, respectively) and at T2 (r=-0.549; r=-0.428 and r=-0.393, respectively). The present study provided evidence that: i) serum IL-23 was up-regulated also in asthmatic children, ii) ICS treatment was able of reducing IL-23, and iii) IL-23 change well related with lung function improvement. Thus, it is presumable that IL-23 could be a suitable marker of allergic inflammation in asthma.

TLR2 and TLR4 gene polymorphisms and atopic dermatitis in Italian children: a multicenter study.

BACKGROUND Genetic factors have an important role in atopic dermatitis (AD) predisposition. Toll like receptor (TLR) are important mediators between environment and immune system. There are incosnsitent studies about TLSR polymorphisms in AD. OBJECTIVE This study examined whether single nucleotide polimorphisms (SNPs) in the genes for TLR2 and TLR4 could be associated with the AD phenotypes and with its clinical severity in a large group of Italian children. METHODS 187 children with Ad and 150 healthy children were recruited. AD severity was assessed by SCORAD. TLR2 (A-16934T and R753Q polymorphisms) and TLR4 (D299G and T399I SNPs) were genotyped by PCR-RFLP. RESULTS The frequency of the R753Q was significantly higher in AD children (16.0 percent) compared with controls (6.0 percent, P =0.004; OR2.99, 95 percent CI 1.39-6.41; RR 1.46, 95 percent CI 1.14-1.69). AD patients a significantly different frequency of the D299G SNP (14.9 percent) in comparison with the controls (6.6 percent, P = 0.01; OR 2.46, 95 percent CI 1.17–5.17; RR 2.24; 95 percent CI 1.15-4.45). CONCLUSION Children with AD may have a distinct genotype and the TLR-2 R753Q SNP was prevalent in a subset of patients with AD characterized by a more severe clinical picture.

Gene-environment interaction in childhood asthma.

The importance of early life environmental influences on the etiology of asthma is implied by the observed geographic and temporal variation in the prevalence of the disease among children. There is evidence pointing to the role of exposure to allergen, various aspects of diet and hygiene-related factors in the etiology of asthma. There is also evidence that heritable factors influence the impact of hygiene-related exposures on the risk of having asthma. A number of important gene-environment interactions have been identified. These interactions point to the biology of environmental exposures as the involved genetic variation is suggestive of certain underlying mechanisms. Polymorphisms within genes coding for the toll-like receptor-lipopolysaccharide (TLR-LPS) signalling pathway may underlie variations in effects of hygiene-related exposures, including specifically endotoxin, on the risk of developing allergic sensitization and allergic disease. This review presents recent findings illustrating the role of gene-environment interactions in childhood asthma susceptibility.

Genetic basis of non syndromic hypodontia: a DNA investigation performed on three couples of monozygotic twins about PAX9 mutation.

INTRODUCTION: Hypodontia, agenesis of one or of more teeth, is a common developmental dental anomaly. To date, over 200 candidate genes have been demonstrated to be active in tooth development. The genes Pax9 plays an important role in the initial stage of odontogenesis. Mutations of Pax9 are associated with autosomal dominant forms of oligodontia, the agenesis of more than six teeth and occasionally of premolars (MIM 604625) in humans. The aim of the present study was to screen the candidate gene causing the non syndromic hypodontia, with agenesis of upper third molars and upper lateral incisors, in three couples of twins. MATERIALS AND METHODS: Peripheral blood samples taken for routine laboratory investigations were used for genotyping. Total genomic DNA was extracted from the buffy coat of 1 ml of EDTA blood samples using phenol-chloroform and the salting out procedure. RESULTS: The insC mutation (nt793, exon4) was observed in the sequencing results by the use of the primers hPAX9ex4F and hPAX9ex4R. InsC raises a frameshift mutation that introduces a nonsense codon so the mRNA activity results impaired. CONCLUSION: In this work, it is described how the same mutation is responsible for a form of dental agenesis--less severe in the number of missing teeth--leading to hypodontia instead of oligodontia. Therefore, it is possible that mutations of the same gene cause different phenotypes; so we can presume that some modifier genes moderate the effect of the first mutation.

Wolfram syndrome and WFS1 gene.

Wolfram syndrome (WS) (MIM 222300) is a rare multisystem neurodegenerative disorder of autosomal recessive inheritance, also known as DIDMOAD (diabetes insipidus, insulin-deficient diabetes mellitus, optic atrophy and deafness). A Wolfram gene (WFS1) has been mapped to chromosome 4p16.1 which encodes an endoplasmic reticulum (ER) membrane-embedded protein. ER localization suggests that WFS1 protein has physiological functions in membrane trafficking, secretion, processing and/or regulation of ER calcium omeostasis. Disturbances or overloading of these functions induce ER stress responses, including apoptosis. Most WS patients carry mutations in this gene, but some studies provided evidence for genetic heterogeneity, and the genotype-phenotype relationships are not clear. Here we review the data regarding the mechanisms and the mutations of WFS1 gene that relate to WS.

TLR4 and NOD2/CARD15 genetic polymorphisms and their possible role in gastric carcinogenesis.

UNLABELLED: The aim of this study was to evaluate the role of TLR4 and NOD2/CARD15 genes in gastric carcinogenesis. PATIENTS AND METHODS: We investigated the allelic frequencies of TLR4 (D299G and T399I) and NOD2/CARD15 (R702W, G908R, and L1007finsC) SNPs in 87 asymptomatic serologically H. pylori-positive individuals (Group I), in 63 patients with antrum-predominant gastritis (Group II) and in 60 patients with corpus-predominant gastritis or pangastritis (Group III). RESULTS: There was significant difference in allelic frequencies of TLR4 D299G SNP in Group II (p=0.02; OR 2.97) as well as in Group III (p=0.001; OR 4.80). Significant difference of T399I SNP allele frequency was only found in Group III (p=0.009; OR 3.73). The allele frequencies of NOD2/CARD15 G908R and of L1007insC SNP were higher in Group III (p=0.003, OR 5.18; p=0.03; OR 3.66, respectively). CONCLUSION: TLR4 and NOD2/CARD15 genes are associated with high risk Group III patients and, therefore, they appear to play a role in gastric carcinogenesis.

Modifications of nuclear envelope in tumour cells of human gastric carcinomas: an ultrastructural study.

BACKGROUND: Various nuclear envelope derivatives, such as the annulate lamellae, the intranuclear tubules as well as the nuclear projections and pockets may be observed electron microscopically in tumour cells. PATIENTS AND METHODS: In a series of eight gastric adenocarcinomas, ultrastructural features of nuclear envelope changes were analyzed and correlated to the biology of the tumours. RESULTS: Histologically, three tumours were intestinal-type adenocarcinomas and showed annulate lamellae in the cytoplasm of some tumor cells. Five tumors were mixed-type adenocarcinomas, with a solid growth pattern; two of these tumours were characterized by the presence of intranuclear tubules, whereas the remaining three tumours exhibited nuclear pockets and projections. Seven out of eight patients died due to metastatic disease during the follow-up period (median 31 months). CONCLUSION: Ultrastructural evaluation of pleomorphism of the nuclear envelope may be an ancillary method for the pathologist in the study of nuclear grading of gastric carcinomas.

Molecular analysis of the CART gene in overweight and obese Italian children using family-based association methods.

AIM: In our study, we evaluated if CART gene A1475G and DeltaA1457 polymorphisms could be associated with obesity. PATIENTS AND METHODS: We recruited 133 Italian trios from among 103 (50 males and 53 females) overweight children (mean age 10.5 years, range 6-14 years; mean BMI 26.1 +/- 3.2 kg/m(2)), and 30 (16 males and 14 females) obese children (mean age 9.0 years, range 6-11 years; mean BMI 32.3 +/- 2.0 kg/m(2)). We also selected 187 non-obese unrelated controls. RESULTS: The allele frequencies of the A1475G single nucleotide polymorphism (SNP) were significantly higher in overweight children (0.07) than in control children (0.02) (p = 0.03) and control adults (0.02) (p = 0.02). Moreover, the allele frequencies were significantly different between obese children (0.08) and control children (0.02) (p = 0.03), and between obese children (0.08) and control adults (0.02) (p = 0.02). The DeltaA1457 SNP showed no significant association with overweight/obesity. TDT statistic revealed a preferential transmission of the 1475G allele from heterozygous parents to overweight children (p < 0.01) and to obese children (p < 0.05). No statistically significant excess transmission of the DeltaA1457 allele was found. CONCLUSION: Our results supported the hypothesis that inherited variations of the CART gene could influence the development of obesity also in Italian children.

RPGRIP1L mutations are mainly associated with the cerebello-renal phenotype of Joubert syndrome-related disorders.

Joubert syndrome-related disorders (JSRDs) are autosomal recessive pleiotropic conditions sharing a peculiar cerebellar and brainstem malformation known as the 'molar tooth sign' (MTS). Recently, mutations in a novel ciliary gene, RPGRIP1L, have been shown to cause both JSRDs and Meckel-Gruber syndrome. We searched for RPGRIP1L mutations in 120 patients with proven MTS and phenotypes representative of all JSRD clinical subgroups. Two homozygous mutations, the previously reported p.T615P in exon 15 and the novel c.2268_2269delA in exon 16, were detected in 2 of 16 families with cerebello-renal presentation ( approximately 12%). Conversely, no pathogenic changes were found in patients with other JSRD phenotypes, suggesting that RPGRIP1L mutations are largely confined to the cerebello-renal subgroup, while they overall represent a rare cause of JSRD (<2%).

Investigation of the eotaxin gene -426C-->T, -384A-->G and 67G-->a single-nucleotide polymorphisms and atopic dermatitis in Italian children using family-based association methods.

BACKGROUND: Eotaxin plays an important role in atopic dermatitis (AD) as a potent chemoattractant and activator of eosinophils and T-helper 2 lymphocytes. AIM: To investigate whether single-nucleotide polymorphisms of the eotaxin gene are associated with AD, we investigated the genotype and allelic frequencies of -426C-->T, -384A-->G, and 67G-->A SNPs in 130 Italian families. METHODS: In total, 130 children with either the extrinsic allergic or intrinsic nonallergic forms of AD (EAD and IAD) were recruited from 130 families. Genotyping was performed using PCR and restriction fragment length polymorphism analysis. RESULTS: A significant difference was observed in the genotype frequency of the -426C-->T SNP between children with EAD and those with IAD (P = 0.01), and between children with EAD and controls (P = 0.01). The allele frequencies of the -426C-->T SNP were significantly different between children with EAD and those with IAD (P < 0.01), and between children with EAD and controls (P < 0.01). For children with EAD, the genotype frequency of the -426C-->T SNP was no different between the groups with mild, moderate and severe SCORAD (P = NS). No significant association was observed between the -384A-->G and 67G-->A SNPs and the two groups of children with EAD and IAD compared with the control group. In 32 trios selected from 68 EAD families, the transmission disequilibrium test showed a preferential transmission of the -426T allele from the parents to affected offspring (P < 0.01). CONCLUSIONS: Our results suggest that in our group of children with AD, the eotaxin gene may play a crucial role in the development of extrinsic AD, probably with other genetic factors.

Gastric adenomas: relationship between clinicopathological findings, Helicobacter pylori infection, APC mutations and COX-2 expression.

Gastric adenomas are rare neoplastic growths characterized by localized polypoid proliferations of dysplastic epithelium that tend to progress to infiltrating adenocarcinoma. Therefore, the identification of molecular markers that could reliably recognize adenomas at risk of progression is advocated in the clinical management. In this study we investigated, in a series of gastric adenoma specimens from an area at high risk of gastric cancer, the relationship between clinicopathological characteristics of adenoma and Helicobacter pylori infection, APC mutational status, and COX-2 and the down-stream enzyme mPGES1 expression. Helicobacter pylori infection, detected in 24%, and 33% by histology and PCR analyses, respectively, did not show any relationship with growth pattern, localization, size, dysplasia grade and presence of synchronous cancer. Pathogenetic mutations of MCR region (codons 1269-1589) of the APC gene were detected only in one case corresponding to a single, small size, low grade, H. pylori-negative adenoma. The expression of COX-2 largely matched that of mPGES(1). Both were overexpressed in 79% of cases showing a relationship with high-grade dysplasia, size >10 mm and presence of a synchronous carcinoma. In conclusion, COX-2 may play a key role in the development and progression of gastric adenoma and could be an attractive target in the management of gastric adenoma at major risk of cancer development.

Molecular analysis of sequence variants in the Fcepsilon receptor I beta gene and IL-4 gene promoter in Italian atopic families.

BACKGROUND: The genetic variants in the Fcepsilon receptor I beta gene (Glu237Gly) and the T allele of the (C590T) polymorphism of interleukin (IL)-4 gene promoter were reported to be associated with atopy. But the data of the studies in different populations are contrasting with one another. METHODS: A group of 25 Italian nuclear families were studied. In each family at least two allergic subjects were present. The allergic children were 65 and the allergic relatives were 35. One hundred and three nonallergic unrelated controls included outpatiens with no history of atopy. The (C590T) promoter polymorphism of the IL-4 and the genetic variant Glu237Gly of Fcepsilon RI beta genes were analysed by the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: A significant difference was observed in the genotype frequency at codon 237 of the Fcepsilon RI beta gene between allergic children and nonatopic control (P < 0.01) and in the allergic relatives (P < 0.001). In the children, the Glu237Gly polymorphism was also associated with elevated circulating levels of immunoglobulin E. The -590C/T allele of IL-4 promoter gene showed no association with atopy. CONCLUSIONS: In our study, the Glu237Gly polymorphism of the Fcepsilon RI beta gene was associated with atopy. Our results have not pointed out an association between the (C590T) promoter polymorphism of the IL-4 gene and atopy. These data suggest the potential role of the Fc RI beta gene in the development of the allergy.

A gene for familial isolated chronic nail candidiasis maps to chromosome 11p12-q12.1.

Chronic mucocutaneous candidiases (CMC) are a group of rare disorders where an altered immune response against Candida leads to persistent and/or recurrent infections of the skin, nails, and mucous membranes. We analysed a five-generation Italian family with an isolated form of CMC, affecting nails only, in the presence of low serum concentration of intercellular adhesion molecule I (ICAM-1). We excluded linkage to candidate regions on chromosomes 2p (CMC with thyroid disease), 21q22.3 (APECED), and 19q13 (ICAM-1). We then carried out a genome-wide scan and assigned the CMC locus to a 19 cM pericentromeric region on chromosome 11.

Morphological evidence of neutrophil-tumor cell phagocytosis (cannibalism) in human gastric adenocarcinomas.

The phenomenon of neutrophil-tumor cell emperipolesis or phagocytosis has been documented by light microscopy in various human carcinomas, but little is known about the cellular pathological processes and the morphological changes involved. In an attempt to clarify the nature of this phenomenon, the authors' ultrastructural studies on the relationships among neutrophils and tumor cells in human gastric carcinomas are reviewed and analyzed. At the electron microscopy level, apoptotic neutrophils were found within vacuoles of adenocarcinoma cells in 2 cases. They showed either early apoptotic morphology with perinuclear chromatin aggregation but cytoplasm integrity or late apoptotic morphology with uniform, collapsed nucleus and tightly packed cytoplasmic granules. A light microscopy review of 200 cases of resected gastric carcinomas identified 22 cases (11%) that were characterized by neutrophil-tumor cell phagocytosis (cannibalism). TUNEL staining confirmed the presence of apoptotic neutrophils within the cytoplasm of the tumor cells. This study provides light and electron microscopic evidence of apoptotic neutrophils phagocytosed by gastric adenocarcinoma cells. The morphological features of neutrophil-tumor cell phagocytosis (cannibalism) would suggest a particular mechanism of tumor-immune escape in human gastric carcinoma.

Eosinophil-tumor cell interaction in advanced gastric carcinoma: an electron microscopic approach.

BACKGROUND: Tumor-associated tissue eosinophils have been observed in human tumors and experimental tumor models, but their function is poorly understood. MATERIALS AND METHODS: One case of intestinal-type adenocarcinoma of the stomach, mainly infiltrated by eosinophils, is studied by light and electron microscopy, focusing on the relationships between eosinophils and tumor cells and on the nature of tumor cell death. RESULTS: Using light microscopy, eosinophils, single or in clusters, were present both in the stroma and within neoplastic glands. With electron microscopy, tumor cells in intimate contact with eosinophils revealed changes consistent with autophagic cell death such as chromatin condensation in small masses into the nucleus, dilation of the nuclear envelope, and numerous cytoplasmic vacuoles. The adenocarcinoma cells, not contacted by neutrophils, remained morphologically well preserved. CONCLUSIONS: Our ultrastructural study suggests the hypothesis of a direct relationship between eosinophil infiltration and induction of autophagic cell death in gastric adenocarcinoma cells.